Abstract
Key message
We demonstrate a new regulatory mechanism in the jasmonic acid (JA) and salicylic acid (SA) mediated crosstalk in potato defense response, wherein, miR160 target StARF16 (a gene involved in growth and development) binds to the promoter of StNPR1 (a defense gene) and negatively regulates its expression to suppress the SA pathway. Overall, our study establishes the importance of StARF16 in regulation of StNPR1 during JA mediated defense response upon necrotrophic pathogen interaction.
Abstract
Plants employ antagonistic crosstalk between salicylic acid (SA) and jasmonic acid (JA) to effectively defend them from pathogens. During biotrophic pathogen attack, SA pathway activates and suppresses the JA pathway via NONEXPRESSOR OF PATHOGENESIS-RELATED GENES 1 (NPR1). However, upon necrotrophic pathogen attack, how JA-mediated defense response suppresses the SA pathway, is still not well-understood. Recently StARF10 (AUXIN RESPONSE FACTOR), a miR160 target, has been shown to regulate SA and binds to the promoter of StGH3.6 (GRETCHEN HAGEN3), a gene proposed to maintain the balance between the free SA and auxin in plants. In the current study, we investigated the role of StARF16 (a miR160 target) in the regulation of the defense gene StNPR1 in potato upon activation of the JA pathway. We observed that a negative correlation exists between StNPR1 and StARF16 upon infection with the pathogen. The results were further confirmed through the exogenous application of SA and JA. Using yeast one-hybrid assay, we demonstrated that StARF16 binds to the StNPR1 promoter through putative ARF binding sites. Additionally, through protoplast transfection and chromatin immunoprecipitation experiments, we showed that StARF16 could bind to the StNPR1 promoter and regulate its expression. Co-transfection assays using promoter deletion constructs established that ARF binding sites are present in the 2.6 kb sequence upstream to the StNPR1 gene and play a key role in its regulation during infection. In summary, we demonstrate the importance of StARF16 in the regulation of StNPR1, and thus SA pathway, during JA-mediated defense response upon necrotrophic pathogen interaction.
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Abbreviations
- NPR1:
-
Nonexpressor of pathogenesis-related genes 1
- PR1:
-
Pathogenesis-related genes 1
- ARF:
-
Auxin response factor
- JA:
-
Jasmonic acid
- SA:
-
Salicylic acid
- MYC2:
-
Jasmonate insensitive 1
- PDF1.2:
-
Defensin-like protein 16
- WRKY6:
-
WRKY containing transcription factor
- miR:
-
miRNA
- GH:
-
Gretchen Hagen3
- 3-AT:
-
3-amino-1,2,4-triazole
- Hpt:
-
Hours post treatment
- Hpi:
-
Hours post infection
- PCR:
-
Polymerase chain reaction
- qRT-PCR:
-
Quantitative real time–PCR
- GUS:
-
Gene encoding β-glucuronidase
- LUC:
-
Luciferase
- St :
-
Solanum tuberosum
- Y1H:
-
Yeast one-hybrid
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Acknowledgements
We thank Dr. Anil Kumar (Thapar University, Patiala, India) for providing us the netrotrophic pathogen Alterneria solani, Dr. Rita Ulloa (INGEBI, Argentina) for sharing with us Solanum tuberosum L. cv. Désirée plants and Prof. Albertha Walhout (MIT, USA) for providing us yeast strains and constructs. We thank Mr. Nitish Lahigude for his help in plant maintenance in the growth chambers and green house, and all the members of molecular plant biology lab at IISER Pune for useful insights and discussions on the project. We also thank IISER Pune for core funding and facility support.
Funding
IISER Pune’s core funding is greatly appreciated. Authors sincerely thank the financial support received from Department of Biotechnology, Government of India, New Delhi (Grant No: BT/PR23716/AGIII/103/1038/2018 dt14.09.2018).
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HSK, BN and AKB conceptualized and designed all the experiments. HSK and AAK performed the experiments and AKB supervised the project. BN assisted with data collection and analysis. HSK, AB, and AKB wrote the manuscript. All authors approve the final version of the manuscript.
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Kalsi, H.S., Karkhanis, A.A., Natarajan, B. et al. AUXIN RESPONSE FACTOR 16 (StARF16) regulates defense gene StNPR1 upon infection with necrotrophic pathogen in potato. Plant Mol Biol 109, 13–28 (2022). https://doi.org/10.1007/s11103-022-01261-0
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DOI: https://doi.org/10.1007/s11103-022-01261-0